GSM5738193: K303R-DoxE2 [ChIP-Seq]; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
TFs and others
Antigen
ESR1
Cell type
Cell type Class
Breast
Cell type
MDA-MB-453
Primary Tissue
Breast
Site of Extraction
Effusion, Pericardial
Tissue Diagnosis
Carcinoma
Attributes by original data submitter
Sample
source_name
MDA-MB-453
cell line
MDA-MB-453
treatment
treated with doxycycline/E2 for 2 days
chip antibody
Anti-ERa (D8H8), CST Japan, #8644
Sequenced DNA Library
library_name
GSM5738193
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
The cells on two 100 mm plates were fixed and nuclear pellets were sonicated to prepare soluble chromatin lysate. ER alpha-chromatin complex was immunoprecipitated by anti-ER alpha antibody from the lysate and precipitated DNA was recovered by reverse crosslinking. The precipitated DNA was purified with AMPure XP beads (Agencourt) and ChIP-seq library was prepared from 2 ng of the purified DNA using SMARTer ThruPLEX DNA-seq Kit (TAKARA). The library was then purified using AMPure XP beads (Agencourt) again. The concentration of the library was quantified using Fluorometer (Quantus) and KAPA library quantification kit (Roche), and the quality was checked by TapeStation D5000 (Agilent). The pooled libraries with unique index sequences were sequenced using NextSeq550 (Illumina) with pair-end 75 bp reads following the manufacturer's protocols.